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牙鲆卵裂雌核发育纯系的建立及鉴定
齐文山;田永胜;姜静;王磊;张文婷;陈红林;张英平;刘万军;孙德强;陈松林

Establishment and Identification of Mitotic Gynogenesis Japanese Flounder (Paralichthys olivaceus) Pure Family

;Tian Yongsheng; ; ; ; ; ; ; ;
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摘要 

在牙鲆家系选择育种中,为了快速的获得具有优良性状的纯系,本研究利用冷冻保存的鲈鱼(Lateolabrax japonicus)精子和2007年构建的牙鲆(Paralichthys olivaceus) 优良家系为实验材料,进行了牙鲆卵裂雌核纯系的诱导建立、遗传倍性分析及遗传标记检测等相关研究。卵裂雌核发育诱导结果表明,在水温为17 ℃条件下,利用灭活鲈鱼精子受精后58 min,以静水压力为590 kg/cm2处理6 min,获得了(32.30±3.34)%的诱导成功率(P<0.05)。利用流式细胞仪对正常胚胎、雌核发育胚胎和单倍体胚胎进行检测,结果表明,单倍体的相对DNA含量为17.07,卵裂雌核发育二倍体相对DNA含量为27.75,正常二倍体相对DNA含量为25.63。雌核家系和近交家系的生长对比结果表明,两者差异极为显著(P<0.05),生长至6个月,近交家系平均体重可达(48.89±17.01) g,而雌核发育纯系体重为(22.09±6.94) g。利用30对微卫星引物对近交家系和卵裂雌核发育纯系进行鉴定与分析,30个微卫星位点在近交系中均表现为杂合多态基因,24个位点在雌核群体中具有多态性,并对这些位点的等位基因数进行了卡方检验,结果表明,等位基因均符合等位基因的1∶1的遗传分离定律。最后计算得到这两个群体的遗传相似性为0.813 9,遗传距离为0.205 9。研究结果提示,利用冷冻保存的异源精子可成功诱导和建立牙鲆有丝分裂雌核发育系,为构建牙鲆优良纯系提供了理论技术依据。

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齐文山
田永胜
姜静
王磊
张文婷
陈红林
张英平
刘万军
孙德强
陈松林
关键词 牙鲆卵裂雌核发育静水压法倍性分析微卫星    
AbstractIn order to facilitate the selective breeding process of Japanese flounder (Paralichthys olivaceus) and obtain the pure family with desirable traits, we employed the cryo-preserved Sea perch (Lateolabrax japonicus) sperm and a flounder family generated in 2007 as experimental materials in this work, which established flounder pure familys with mitotic gynogenesis, analyzed the ploidy and monitored genetic markers. The results from mitotic gynogenesis suggested: In 17 ℃ water, inactivating sea perch sperm in 58 min after fertilization, followed by 6 min of hydrostatic pressure (590 kg/cm2), led to a higher rate of induction (32.30±3.34)%. Detection on three types of embryos (haploid, mitotic gynogenesis and normal diploid) by flow Cytometer showed that the relative amounts of DNA were 17.07 (mode: 17) in the haploid, 27.75 in mitotic gynogenesis (mode: 27) and 25.63 (mode: 27) in normal diploid. When the two familys (mitotic gynogenesis family and inbreeding family) were compared, their growth difference was significant. The average weight of inbreeding group was about (48.89±17.01) g at 6-month fish, while the weight of gynogenetic family was (22.09±6.94) g. Thirty microsatellite markers were used to analyze the inbreeding and gynogenetic familys, the data indicated that all microsatellite sites showed heterozygous gene polymorphism in inbreeding group and 24 loci were with polymorphism in the mitotic gynogenesis group. In parallel, the chi-square test was used and the results implied that the segregation rate was in accordance with the 1∶1 (alleles). The genetic similarity between inbreed group and gynogenesis group was 0.813 9, while the genetic distance was 0.205 9. It was concluded that mitotic gynogenesis familys can be generated via usage of cryo-preserved sea perch sperm, which provides the theoretical basis for establishing superior Japanese flounder familys.
Key wordsParalichthys olivaceus    Mitotic gynogenesis    Hydraulic pressure    Ploidy investigation    Microsatellites
收稿日期: 1900-01-01     
引用本文:   
齐文山;田永胜;姜静;王磊;张文婷;陈红林;张英平;刘万军;孙德强;陈松林. 牙鲆卵裂雌核发育纯系的建立及鉴定[J]. , 2014, 22(5): 541-551.
;Tian Yongsheng; ; ; ; ; ; ; ; .

Establishment and Identification of Mitotic Gynogenesis Japanese Flounder (Paralichthys olivaceus) Pure Family

. , 2014, 22(5): 541-551.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/      或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2014/V22/I5/541
 
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